Abstract Prenatal alcohol exposure induces neuronal death in the fetal brain through apoptotic mechanism. Evidence shows that alcohol exposure may accelerate apoptosis in the developing brain through direct activation of mitochondrial apoptotic signaling pathways. Although studies have demonstrated that prenatal alcohol exposure induces mitochondrial dysfunction, relatively little is known about the effects of alcohol exposure on mitochondrial signaling pathways. In fact, most studies have focused on the effects of alcohol exposure on mitochondrial anatomical changes and enzymes activities. In contrast, we will use state-of-the-art proteomics technology, microarray, and other assays to investigate the effects of prenatal alcohol exposure on mitochondrial signaling pathways. In addition, studies conducted at NIH-NICHD and Tel Aviv University have suggested that two peptides play a key role in protecting mitochondria against oxidative stress including alcohol. One is a nine amino acid peptide (SALLRSIPA), known as SAL or ADNF-9, derived from activity- dependent neurotrophic factor (ADNF);the other is an eight amino acid peptide (NAPVSIPQ), known as NAP, derived from activity-dependent neuroprotective protein (ADNP). In a model of fetal alcohol syndrome (FAS), these peptides have been shown to prevent alcohol-induced decreases in reduced mitochondrial glutathione. In the present proposal, we will use our established liquid diet mixed with alcohol in a model of fetal alcohol exposure (FAE) that mimics moderate alcohol drinking. Because the molecular mechanisms of action of both SAL and NAP in protecting mitochondria against alcohol exposure are not fully characterized, we aim to delineate the precise mitochondrial signaling pathways that mediate SAL/NAP protection against prenatal alcohol exposure in the fetal brain. Based on in vitro and in vivo studies on mitochondria related to oxidative stress, we hypothesize that: 1) Prenatal alcohol exposure induces mitochondrial dysfunction and subsequently cell death through mitochondrial signaling pathways involving Bcl-2 family;2) Prenatal alcohol exposure alters nuclear and mitochondrial encoded proteins to subsequently induce apoptosis;3) ADNF-9 and/or NAP protect directly and/or indirectly mitochondrial dysfunction against the insult of prenatal alcohol exposure. The data generated from this application will provide a detailed understanding of the effects of prenatal alcohol exposure on mitochondrial signaling pathways and the role of ADNF-9 and NAP in protecting mitochondria in a model of FAE. Public Health Relevance: This study will investigate the effect or alcohol exposure during pregnancy on an organelle termed "mitochondria'which play a respiratory role in cells. We will also investigate the role of molecules that have a protective effect against alcohol exposure on "mitochondria" of the central nervous system during pregnancy stages. This will provide information in finding therapeutic ways for the treatment of fetal alcohol exposure.